Comparison of Modified Papanicolaou and Dane-Herman Stain with Routine Hematoxylin and Eosin Stain for Demonstration of Keratin in Tissue Sections
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Date
2019-04
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Addis Ababa University
Abstract
Background: Keratins are intermediate filament forming proteins with specific physicochemical properties produced in any vertebrate epithelia. Having characteristic findings in many epithelial pathologies made them diagnostically important marker. Hematoxylin and Eosin(HE) stain, the routine dye, does not distinguish different eosinophilic components of connective tissue distinctly. On the other hand, special stains like modified Papanicolaou( Mod Pap) and Dane-Herman(D-H) stain can prove to be a simple solution to detect the presence and pattern of keratin histologically.
Objectives: To compare modified Papanicolaou stain and Dane Herman stain with routine Hematoxylin and Eosin Stain for demonstration of keratin and also there basic staining characteristics on paraffin blocks tissue sections.
Methodology: A total of 116 Formalin-Fixed Paraffin-Embedded(FFPE) tissue blocks were taken of known keratin containing tissue which included Squamous Cell Carcinoma(SCC), Squamous Papilloma, Verrucous Carcinoma and Odontogenic Keratocyst. The tissue blocks were retrieved from the archive of Tikur Anbessa Specialized Hospital(TASH), Department of Pathology. Three
sections of 4µm thick tissue from each paraffin block were made and stained with HE, Mod PAP and D-H stains. Then the three stains were evaluated for Nuclear stain(NS), Cytoplasmic stain(CS), Uniformity of staining(US), Background stain(BS) and Keratin pattern(KP) by two pathologists. Finally, the third Senior pathologist was used as a tie breaker.
Result: Mod PAP showed agreement with HE by showing Mc Nemar’s P value of 0.219 for NS, 0.727 for CS, 0.851 for US, 0.824 for BS and 1.00 for KP. Whereas D-H showed disagreement with HE for the first four parameters(P=0.000). But D-H showed agreement for KP. Thus, these
results show that keratin was stained by D-H, Mod PAP and HE stains distinctly. Mod PAP stain showed similar result when compared with HE stain with parameters-NS, CS, US and BS. However, D-H stain showed statistically significant difference when crosschecked with HE stains. Special stains agreement were analyzed using McNemar as a statistical data tool, P< 0.05 were considered statistically significant.
Conclusion: Mod PAP and D-H stains showed similar result for staining of keratin compared to HE stain. For the other four parameters Mod PAP showed an excellent result but not D-H.
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Words: Keratin, Dane-Herman, Modified Papanicolaou, Hematoxylin and Eosin stain